荧光标记MOMP抗体检测沙眼衣原体的方法及性能评价
|
摘要
目的建立异硫氰酸荧光素标记沙眼衣原体主要外膜蛋白单克隆抗体直接免疫荧光法检测沙眼衣原体的方法 ,并评价其灵敏度和特异性。方法双盲法。采集女性宫颈拭子双份,一份免疫荧光法检测,另一份采用荧光定量PCR检测。以PCR测定结果作为标准,分别计算DFA和PCR结果的一致性。结果共检测142份宫颈拭子,准确率为99.3%,其中阳性符合率为90.9%,阴性符合率为100%。Kappa系数一致性检验,k为0. 894。荧光标记MOMP抗体检测沙眼衣原体灵敏度、特异性与PCR两种方法检测结果一致。结论异硫氰酸标记沙眼衣原体主要外膜蛋白单克隆抗体法检测沙眼衣原体有较高的灵敏度和特异性,可用于临床检验。
Objective Chlamydia trachomatis is considered as the most common causing factor of sexually transmitted diseases. A test method of fluorescein isothiocyanate(FITC) labeled major outer membrane protein(MOMP) assay for the detection of Chlamydia trachomatis antigen was established in this study. The sensitivity and specificity of the test were also evaluated. Methods The double endocervical swabs from female patients were collected following the standard procedures. One sample was detected for Chlamydia trachomatis by using PCR through the amplification of a specific sequence and the other sample was detected for Chlamydia trachomatis by using FITC labeled MOMP assay. The double blind method was used to test the consistence. Results Out of 142 samples tested,the correct rate of FITC labeled MOMP assay was 99. 3%.The sensitivity and specificity of the assay was 90. 9% and 100%,respectively. Kappa value was 0. 894.Conclusion FITC labeled MOMP assay for the detection of Chlamydia trachomatis has a high sensitivity and specificity and could be used in the clinical detection of Chlamydia trachomatis.
Objective Chlamydia trachomatis is considered as the most common causing factor of sexually transmitted diseases. A test method of fluorescein isothiocyanate(FITC) labeled major outer membrane protein(MOMP) assay for the detection of Chlamydia trachomatis antigen was established in this study. The sensitivity and specificity of the test were also evaluated. Methods The double endocervical swabs from female patients were collected following the standard procedures. One sample was detected for Chlamydia trachomatis by using PCR through the amplification of a specific sequence and the other sample was detected for Chlamydia trachomatis by using FITC labeled MOMP assay. The double blind method was used to test the consistence. Results Out of 142 samples tested,the correct rate of FITC labeled MOMP assay was 99. 3%.The sensitivity and specificity of the assay was 90. 9% and 100%,respectively. Kappa value was 0. 894.Conclusion FITC labeled MOMP assay for the detection of Chlamydia trachomatis has a high sensitivity and specificity and could be used in the clinical detection of Chlamydia trachomatis.
引文
[1]倪安平,沈浣.微量沙眼衣原体组织培养法的建立及临床应用[J].生殖医学杂志,1998,7(1):10-13.
[2]李永哲,倪安平,汪波,等.应用免疫荧光检测沙眼衣原体[J].中华医学检验杂志,1995,18(1):29-31.
[3]闫玲,冯树异,周劲松,等.应用McCoy细胞及Hep-2细胞培养沙眼衣原体D型株[J].北京医科大学学报,1997,29(1):83-83.
[4]倪安平,朱晓春,王宝玺,等.全自动荧光酶免疫分析仪检测泌尿生殖道衣原体的评价[J].中华检验医学杂志,2001,24(3):190-192.
[5]XUE Y, ZHENG H, TANG W, et al. Prevalence and genotype distribution of Chlamydia trachomatis inurine among men attending sexually transmitted disease clinics in Guangdong Province,China,in 2016[J].Jpn J Infect Dis,2018,71(2):104-108.
[6]韦红,吴仕孝,余加林,等.连接酶链反应-酶联免疫吸附试验诊断沙眼衣原体感染的初步应用[J].中华检验医学杂志,2004,27(5):295-298.
[7]刘小平,樊尚荣,周惠平,等.不同人群妇女宫颈沙眼衣原体感染的检测[J].中华医院管理杂志,1995,11(12):729-730.
[8]刘小平,樊尚荣,李建武,等.深圳市人工流产女青年生殖道感染状况及相关因素分析[J].中国全科医学杂志,2004,7(15):1052-1054.
[9]IMUDIA A N, DETTI I, PUSCHCK E E, et al. The prevalence of urea plasma urealyticum, myco plasma hominis, chlamydia trachomatis and neisseria gonorrhoeae infections, and the rubella status of patients undergoing an initial infertility evaluation[J]. J Assist Reproduction Genet,2008,25(1):43-46.
[10]周淑群,书柳华,周定球,等.支原体、衣原体感染与不孕症的关系及耐药性分析[J].中华医院感染学杂志,2009,19(10):1314-1316.
[11]徐三兰,云国家.PCR检测解脲原体和沙眼衣原体感染与女性不孕关系分析[J].中国优生与遗传杂志,2010,18(2):120-122
[12]TOYOKAWA M, KISHIMOTO T, CAI Y, et al. Severe Chlamydophila psittaci pneumonia rapidly diagnosed by detection of antigen in sputum with an immunochromatography assay[J]. J Infect Chemother,2004,10(4):245-249.
[13]HE W, FELDERMAN M, EVANS A C, et al. Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein(MOMP)for vaccine development[J]. J Biol Chem,2017,292(36):15121-15132.
[14]TIFREA D F,RALLI-JAIN P, PAL S, et al. Vaccination with the recombinant major outer membrane protein elicits antibodies to the constant domains and induces cross-serovar protection against intranasal challenge with chlamydia trachomatis[J]. Infect Immun,2013,81(5):1741-1750.
[15]WEN Z,BODDICKER M A,KAUFHOLD R M,et al. Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research[J].BMC Microbiol,2016,16(1):1-13.
[16]樊尚荣,李健玲(编译).2010年美国疾病控制中心沙眼衣原体感染治疗指南[J].中国全科医学,2011,14(2B):464-465.
[17]KHAN E R,HOSSAIN M A,PAUL S K,et al. Comparison between ICT and PCR for diagnosis of Chlamydia trachomatis[J].Mymensingh Med J,2012,21(2):190-194.
[2]李永哲,倪安平,汪波,等.应用免疫荧光检测沙眼衣原体[J].中华医学检验杂志,1995,18(1):29-31.
[3]闫玲,冯树异,周劲松,等.应用McCoy细胞及Hep-2细胞培养沙眼衣原体D型株[J].北京医科大学学报,1997,29(1):83-83.
[4]倪安平,朱晓春,王宝玺,等.全自动荧光酶免疫分析仪检测泌尿生殖道衣原体的评价[J].中华检验医学杂志,2001,24(3):190-192.
[5]XUE Y, ZHENG H, TANG W, et al. Prevalence and genotype distribution of Chlamydia trachomatis inurine among men attending sexually transmitted disease clinics in Guangdong Province,China,in 2016[J].Jpn J Infect Dis,2018,71(2):104-108.
[6]韦红,吴仕孝,余加林,等.连接酶链反应-酶联免疫吸附试验诊断沙眼衣原体感染的初步应用[J].中华检验医学杂志,2004,27(5):295-298.
[7]刘小平,樊尚荣,周惠平,等.不同人群妇女宫颈沙眼衣原体感染的检测[J].中华医院管理杂志,1995,11(12):729-730.
[8]刘小平,樊尚荣,李建武,等.深圳市人工流产女青年生殖道感染状况及相关因素分析[J].中国全科医学杂志,2004,7(15):1052-1054.
[9]IMUDIA A N, DETTI I, PUSCHCK E E, et al. The prevalence of urea plasma urealyticum, myco plasma hominis, chlamydia trachomatis and neisseria gonorrhoeae infections, and the rubella status of patients undergoing an initial infertility evaluation[J]. J Assist Reproduction Genet,2008,25(1):43-46.
[10]周淑群,书柳华,周定球,等.支原体、衣原体感染与不孕症的关系及耐药性分析[J].中华医院感染学杂志,2009,19(10):1314-1316.
[11]徐三兰,云国家.PCR检测解脲原体和沙眼衣原体感染与女性不孕关系分析[J].中国优生与遗传杂志,2010,18(2):120-122
[12]TOYOKAWA M, KISHIMOTO T, CAI Y, et al. Severe Chlamydophila psittaci pneumonia rapidly diagnosed by detection of antigen in sputum with an immunochromatography assay[J]. J Infect Chemother,2004,10(4):245-249.
[13]HE W, FELDERMAN M, EVANS A C, et al. Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein(MOMP)for vaccine development[J]. J Biol Chem,2017,292(36):15121-15132.
[14]TIFREA D F,RALLI-JAIN P, PAL S, et al. Vaccination with the recombinant major outer membrane protein elicits antibodies to the constant domains and induces cross-serovar protection against intranasal challenge with chlamydia trachomatis[J]. Infect Immun,2013,81(5):1741-1750.
[15]WEN Z,BODDICKER M A,KAUFHOLD R M,et al. Recombinant expression of Chlamydia trachomatis major outer membrane protein in E. Coli outer membrane as a substrate for vaccine research[J].BMC Microbiol,2016,16(1):1-13.
[16]樊尚荣,李健玲(编译).2010年美国疾病控制中心沙眼衣原体感染治疗指南[J].中国全科医学,2011,14(2B):464-465.
[17]KHAN E R,HOSSAIN M A,PAUL S K,et al. Comparison between ICT and PCR for diagnosis of Chlamydia trachomatis[J].Mymensingh Med J,2012,21(2):190-194.